Methods of treating multifocal cancer

ABSTRACT

The embodiments include methods of treating (preventing or reducing the incidence of) multifocal cancer by administering to a unifocal cancer focus a composition comprising a therapeutically effective amount of a therapeutically effective amount of pharmaceutically active ingredient capable of inducing necrosis of the unifocal cancer tumor, wherein administration reduces multifocal cancer incidence, multifocal cancer grade, and multifocal cancer progression (worsening) in the entire organ or organism.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted electronically in ASCII format and is hereby incorporated byreference in its entirety. Said ASCII copy, created on Jul. 31, 2019, isnamed NYMOX-0505482_ST25.txt and is 480 bytes in size.

BACKGROUND 1. Field of the Embodiments

The embodiments include methods of treating multifocal cancer in mammalshaving multifocal cancer, and more specifically to methods of preventingand/or reducing multi-focal cancer development and progression in theentire affected organ or organism by directly administering to a single(unifocal) cancer tumor a composition comprising a pharmaceuticallyactive ingredient capable of inducing necrosis of the tumor and apharmaceutically acceptable carrier. In one embodiment, thepharmaceutically active ingredient is fexapotide triflutate (“FT”), andthe multifocal cancer is prostate cancer. The methods include, but arenot limited to, administering the compositions intramuscularly, orally,intravenously, intraperitoneally, intraprostatically, intracerebrally(intraparenchymally), intracerebroventricularly, intralesionally,intraocularly, intraarterially, intrathecally, intratumorally,intranasally, topically, transdermally, subcutaneously, or intradermallyto patients in need thereof, wherein targeted administration of thecompositions to the unifocal cancer tumor reduces or prevents multifocalcancer incidence, multifocal cancer grade, and multifocal cancerprogression (worsening).

2. Description of Related Art

The essence of many medical treatments and procedures involves theremoval or destruction of harmful or unwanted tissue. Examples of suchtreatments include the surgical removal of cancerous or pre-cancerousgrowths, the destruction of metatastic tumors through chemotherapy, andthe reduction of glandular (e.g. prostate) hyperplasia. Other examplesinclude the removal of unwanted facial hair, the removal of warts, andthe removal of unwanted fatty tissue.

There is a need for an effective composition that will destroy and henceeither facilitate the removal of or inhibit the further growth ofharmful or unwanted cells and tissue but will have mainly local effectsand minimal or absent systemic toxicity. There also is a need to reducethe need for invasive surgical intervention, radiation and/orchemotherapy, and other invasive procedures.

Some agents known to have the ability to destroy and hence eitherfacilitate the removal of or inhibit the further growth of harmful orunwanted cells and tissue are disclosed in U.S. patent application Ser.No. 14/808,713, filed Jul. 24, 2015, entitled: METHODS OF REDUCING THENEED FOR SURGERY IN PATIENTS SUFFERING FROM BENIGN PROSTATICHYPERPLASIA; U.S. patent application Ser. No. 14/606,683, filed Jan. 27,2015, entitled: METHOD OF TREATING DISORDERS REQUIRING DESTRUCTION ORREMOVAL OF CELLS, U.S. application Ser. No. 14/738,551, filed Jun. 12,2015, entitled: COMBINATION COMPOSITIONS FOR TREATING DISORDERSREQUIRING REMOVAL OR DESTRUCTION OF UNWANTED CELLULAR PROLIFERATIONS,U.S. patent application Publication Nos. 2007/0237780 (now abandoned);2003/0054990 (now U.S. Pat. No. 7,172,893); 2003/0096350 (now U.S. Pat.No. 6,924,266); 2003/0096756 (now U.S. Pat. No. 7,192,929); 2003/0109437(now U.S. Pat. No. 7,241,738); 2003/0166569 (now U.S. Pat. No.7,317,077); 2005/0032704 (now U.S. Pat. No. 7,408,021); and 2015/0148303(now U.S. Pat. No. 9,243,035), the disclosures of each of which areincorporated by reference herein in their entirety.

One of the agents disclosed in these documents is fexapotide triflutate,or FT. FT has been shown to reduce prostate glandular cells, toameliorate or reduce LUTS, and to treat BPH in men with prostateenlargement. FT also has been disclosed as useful in reducing the onsetof prostate cancer by treating BPH in a mammal having BPH in which thecompositions containing FT are administered to the mammal in transitionzone (central) prostate. See, e.g., U.S. Pat. No. 10,183,058, thedisclosure of which is incorporated by reference herein in its entirety.FT also is the subject of a clinical trial to assess the efficacy of FTin treating a low grade low risk localized (T1c) prostate cancer tumor,in which the trial is designed to assess the efficacy of FT ineliminating the localized tumor within 45 days post-treatment.

Prostate cancer is known as a disease with an extremely high prevalencerelative to its clinical incidence in the population. Prostate cancerhas a high asymptomatic incidence and a long asymptomatic duration.Prostate cancer has an interval of 7 to 14 years on average during whichthe cancer is present but is preclinical because it is not detected bytypical clinical or laboratory examinations (see Etzioni, R et al., Am JEpidemiol. Vol. 148, pp. 775-85 (1998); and Gulati, R, et al., CancerEpidemiol Biomarkers Prev; Vol. 20(5), pp. 740-50 (2011)).

Prostate cancer is often considered a multifocal disorder in which theprostate gland includes multiple adenocarcinoma foci of varyingheterogeneity. This makes the cancer difficult to treat effectively,often resulting in radical prostatectomy, which causes numerous lifealtering issues for men including erectile dysfunction and urinaryincontinence. Some prostate cancer, however, is considered low grade lowrisk localized (T1c) prostate cancer, which has been reported in fromabout 20 to about 35% of radical prostatectomy specimens. Mazzucchelli,et al., “Pathology of Prostate Cancer and Focal Therapy (MaleLumpectomy'),” Anticancer Research, Vol. 29, pp. 5155-5162 (2009);Ibeawuchi, et al., “Genome-Wide Investigation of Multifocal and UnifocalProstate Cancer—Are They Genetically Different?” Int. J. Mol. Sci., Vol.14, ppp. 11816-11829 (2013).

Due to the severity of radical prostatectomy, recent studies havereported on focal therapy in which a portion of the prostate ispreserved, although the efficacy of focal therapy of prostate cancer andpreventing cancer progression remains uncertain. Quann, et al., “Currentprostate biopsy protocols cannot reliably identify patients for focaltherapy; . . . ”, Int. J. Clin. Exp. Pathol., Vol. 3(4), pp. 401-407(2010). Identifying, targeting, and focally destroying a specific tumorhas yet to be realized (Mazzucchelli at 5159), and to date focal therapyinvolves ablation of large portions, (e.g., an entire lobe, orhemiablation), of the prostate. It therefore was not heretofore known orexpected that treating a low grade low risk unifocal prostate cancertumor by targeting just the unifocal tumor would be effective inreducing cancer incidence, cancer grade, and cancer progression(worsening) throughout the entire prostate. As a consequence, theclinical trials in which FT was assessed to treat only the unifocalprostate cancer was not designed, and not expected to be effective intreating the entire prostate.

A common technique used in identifying and monitoring prostate cancer isassessing PSA levels in conjunction with biopsy evaluation. Typicalbiopsy of the prostate usually involves taking numerous samples throughthe prostate gland, and assessing the tissue using a Gleason Score. TheGleason Score measures how abnormal a cancer cell looks under amicroscope and is a good indicator of how quickly the cells are likelyto grow and spread. The Gleason Score is calculated by adding togetherthe two grades of cancer that make up the largest areas of the biopsiedtissue sample, and is often represented as two numbers, such as 3+3,along with the total score, such as 6. The primary pattern Gleason scoreis listed first and the secondary second. A biopsy having a Gleasongrade of 3+4 (total 7) is considered less severe than a biopsy having aGleason grade of 4+3 (total 7), because the primary pattern is above 3.Accordingly, clinicians review not only the total Gleason score, but thevalues of the primary and secondary pattern when assessing the severityof the tumor(s). The following table provides a classification of thegroups of prostate cancer and the associated risks:

Risk Group ISUP Grade Group Gleason Score Low Grade Group 1 GleasonScore ≤6 Intermediate Favorable Grade Group 2 Gleason Score 7 (3 + 4)Intermediate Unfavorable Grade Group 3 Gleason Score 7 (4 + 3) HighGrade Group 4 Gleason Score 8 High Grade Group 5 Gleason Score 9-10

For low grade low risk localized (T1c) prostate cancer in which theGleason Score is ≤6. Active Surveillance (AS) is the ordinary course oftreatment. This is because, while some may very well mature to high riskcancer requiring radical prostatectomy (e.g., Gleason Score ≥8), many donot. As noted above, up to about 35% of prostates surgically removedonly have low grade low risk localized (T1c) prostate cancer.Accordingly, those patients who had their prostates surgically removedbut still did not have high risk cancerous tumors needlessly sufferedthe deleterious effects of radical prostatectomy surgery. Accordingly,the typical standard of care is that, when the Gleason Score is 7 orabove, especially when the primary pattern is 4, corrective measures aretaken, either by removing a large portion of the prostate, chemotherapyor radiation, or a radical prostatectomy. It therefore would bedesirable to discover a safe and effective manner of treating prostatecancer patients in the low and/or low and intermediate risk groups byfocal treatment of a single foci, in which case the treatment iseffective in ameliorating, reducing, and/or preventing the progressionof the cancer throughout the entire prostate.

Like prostate cancer, other cancers are known to be multifocal innature, and preceded by and associated with unifocal tumors, or small,cancerous tumors that are not significant enough to warrant treatment.Such multifocal cancers include, but are not limited to one or more ofthe following: squamous cell head and neck carcinoma (HNSCC), cutaneoussquamous cell carcinoma (skin SCC), breast cancer, adenocarcinoma andSCC of lung, esopahgeal cancer, gastric cancer, colon cancer, bladdercancer, cervical cancer, melanoma, brain cancer pancreatic cancer,ovarian cancer, bone marrow cancer, and leukemia.

Throughout this description, including the foregoing description ofrelated art, any and all publicly available documents described herein,including any and all U.S. patent published patent applications, arespecifically incorporated by reference herein in their entirety. Theforegoing description of related art is not intended in any way as anadmission that any of the documents described therein, including pendingU.S. patent applications, are prior art to the present disclosure.Moreover, the description herein of any disadvantages associated withthe described products, methods, and/or apparatus, is not intended tolimit the embodiments. Indeed, aspects of the embodiments may includecertain features of the described products, methods, and/or apparatuswithout suffering from their described disadvantages.

SUMMARY OF THE EMBODIMENTS

There remains a need in the art for new, less toxic, and less frequent,and essentially non-invasive treatments for preventing or reducing theprogression or incidence of multifocal cancer. There also remains a needin the art for such treatments that reduce the incidence of multifocalcancers. The embodiments satisfy these needs.

This disclosure is premised in part on the discovery thatpharmaceutically active ingredients capable of inducing necrosis of aunifocal cancer tumor are capable of being administered to the tumor,but yet have an unexpected effect of reducing multifocal cancerincidence, reducing multifocal cancer grade, and reducing multifocalprogression (worsening) in the entire affected organ or organism.Suitable pharmaceutically active ingredients capable of inducingnecrosis of such tumors, include, for example, fexapotide triflutate,(FT), a peptide described by the amino acid sequenceIle-Asp-Gln-Gln-Val-Leu-Ser-Arg-Ile-Lys-Leu-Glu-Ile-Lys-Arg-Cys-Leu (SEQID NO. 1), ZYTIGA® (abiraterone acetate), EARLEADA® (apalutamide),abazitaxel, CASODEX® (Bicalutamide), ELIGARD® and LUPRON®, (LeuprolideAcetate), FIRMAGON® (Degarelix), flutamide, JEVTANA® (Cabazitaxel),mitoxantrone hydrochloride, NILANDRON® (Nilutamide), PROVENGE®(Sipuleucel-T), TAXOTERE® (Docetaxel), XOFIGO® (Radium 223 Dichloride),XTANDI® (Enzalutamide), ZOLADEX® (Goserelin Acetate), and mixtures andcombinations thereof. Such administration was unexpectedly found, afterseveral years of ongoing follow up, to be effective in reducingmultifocal cancer incidence, reducing multifocal cancer grade, andreducing multifocal progression (worsening) in the entire affected organor organism and was not limited to local efficacy. The embodimentstherefore are capable of vastly improving the quality of life of manymen suffering from multifocal cancer that would have otherwise undergonea more aggressive treatment, like ablation of a large portion of theorgan, chemotherapy, radiation, or complete removal of the gland (e.g.,radical prostatectomy, colonectomy, lung removal or transplantation, andthe like.

Some embodiments are directed to methods of reducing multifocal cancerincidence, reducing multifocal cancer grade, and reducing multifocalcancer progression (worsening) in mammals having low grades or at lowrisk of multifocal cancer (i.e., for prostate cancer, having a GleasonScore ≤6) by administering to the mammal a therapeutically effectiveamount of a composition comprising at least one pharmaceutically activeingredient capable of inducing necrosis of the low grade, low risk,localized cancer tumor. The method includes administering atherapeutically effective amount of the composition to a singlecancerous foci (unifocal tumor), and reducing the percentage of mammalsexhibiting 1 new focus in the entire organ or organism.

In certain embodiments, the method includes administering atherapeutically effective amount of FT to a single cancerous foci(unifocal tumor) in the prostate of the mammal, and reducing thepercentage of mammals exhibiting ≥1 new focus with an increase inGleason grade (new multifocal with Gleason upgrade) in the entireprostate by an amount of from about 40% to about 100%, when compared toactive surveillance, when measured at least 18 months after treatment,or from about 30% to about 90%, when compared to active surveillance,when measured at least 36 months after treatment, or from about 5% toabout 85%, when compared to active surveillance, when measured at least48 months after treatment. The methods also include administering atherapeutically effective amount of the composition to a singlecancerous foci (unifocal tumor) in the prostate of the mammal, andreducing the percentage of mammals exhibiting ≥1 new focus in the entireprostate with an increase in Gleason grade primary pattern (newmultifocal with Gleason primary pattern ≥4) by an amount of from about40% to about 100%, when compared to active surveillance, when measuredat least 18 months after treatment, or from about 30% to about 100%,when compared to active surveillance, when measured at least 36 monthsafter treatment, or from about 20% to about 100%, when compared toactive surveillance, when measured at least 48 months after treatment.

The compositions can be administered intramuscularly, orally,intravenously, intraperitoneally, intracerebrally (intraparenchymally),intracerebroventricularly, intratumorally, intralesionally,intradermally, intrathecally, intranasally, intraocularly,intraarterially, topically, transdermally, via an aerosol, infusion,bolus injection, implantation device, sustained release system etc.

In another embodiment, the composition includes a therapeuticallyeffective amount of FT that is administered in an amount ranging fromabout 2.0 mg to about 20 mg.

In another embodiment, the method includes administering atherapeutically effective amount of FT to a single cancerous foci(unifocal tumor) in the prostate of the mammal, and reducing thepercentage of mammals having conventional cancer treatment (surgery,radiotherapy, or chemotherapy), and exhibiting ≥1 new focus in theentire prostate with an increase in Gleason grade (new multifocal withGleason upgrade) by an amount of from about 50% to about 100%, whencompared to active surveillance, when measured at least 18 months aftertreatment, or from about 40% to about 90%, when compared to activesurveillance, when measured at least 36 months after treatment, or fromabout 35% to about 85%, when compared to active surveillance, whenmeasured at least 48 months after treatment. The methods also includeadministering a therapeutically effective amount of FT to a singlecancerous foci (unifocal tumor) in the prostate of the mammal, andreducing the percentage of mammals having conventional cancer treatment(surgery, radiotherapy, or chemotherapy), and exhibiting ≥1 new focus inthe entire prostate with an increase in Gleason grade primary pattern(new multifocal with Gleason primary pattern ≥4) by an amount of fromabout 70% to about 100%, when compared to active surveillance, whenmeasured at least 18 months after treatment, or from about 60% to about100%, when compared to active surveillance, when measured at least 36months after treatment, or from about 50% to about 100%, when comparedto active surveillance, when measured at least 48 months aftertreatment.

Both the foregoing general description and the following detaileddescription are exemplary and explanatory and are intended to providefurther explanation of the embodiments as claimed. Other objects,advantages, and features will be readily apparent to those skilled inthe art from the following detailed description of the embodiments.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

Before the present proteins, nucleotide sequences, peptides,compositions, active agents, etc., and methods are described, it isunderstood that this invention is not limited to the particularmethodology, protocols, cell lines, vectors, and reagents described, asthese may vary. It also is to be understood that the terminology usedherein is for the purpose of describing particular embodiments only, andis not intended to limit the scope of the present embodiments which willbe limited only by the appended claims.

Terms and phrases used herein are defined as set forth below unlessotherwise specified. Throughout this description, the singular forms“a,” “an,” and “the” include plural reference unless the context clearlydictates otherwise. Thus, for example, a reference to “a host cell”includes a plurality of such host cells, and a reference to “anantibody” is a reference to one or more antibodies and equivalentsthereof known to those skilled in the art, and so forth.

Amino acids and amino acid residues described herein may be referred toaccording to the accepted one or three-letter code provided in the tablebelow.

TABLE 1 Three-Letter Amino Acid One-Letter Symbol Symbol Alanine A AlaArginine R Arg Asparagine N Asn Aspartic acid D Asp Cysteine C CysGlutamine Q Gln Glutamic acid E Glu Glycine G Gly Histidine H HisIsoleucine I Ile Leucine L Leu Lysine K Lys Methionine M MetPhenylalanine F Phe Proline P Pro Serine S Ser Threonine T ThrTryptophan W Trp Tyrosine Y Tyr Valine V Val

The expression “pharmaceutically active ingredients capable of inducingnecrosis of such tumors” denotes, for example, fexapotide triflutate,(FT), a peptide described by the amino acid sequenceIle-Asp-Gln-Gln-Val-Leu-Ser-Arg-Ile-Lys-Leu-Glu-Ile-Lys-Arg-Cys-Leu (SEQID NO. 1), ZYTIGA® (abiraterone acetate), EARLEADA® (apalutamide),abazitaxel, CASODEX® (Bicalutamide), ELIGARD® and LUPRON®, (LeuprolideAcetate), FIRMAGON® (Degarelix), flutamide, JEVTANA® (Cabazitaxel),mitoxantrone hydrochloride, NILANDRON® (Nilutamide), PROVENGE®(Sipuleucel-T), TAXOTERE® (Docetaxel), XOFIGO® (Radium 223 Dichloride),XTANDI® (Enzalutamide), ZOLADEX® (Goserelin Acetate), and mixtures andcombinations thereof. Fexapotide Triflutate (“FT”), as it is usedherein, denotes a 17-mer peptide having the amino acid sequence:Ile-Asp-Gln-Gln-Val-Leu-Ser-Arg-Ile-Lys-Leu-Glu-Ile-Lys-Arg-Cys-Leu (SEQID NO. 1). FT is disclosed in U.S. Pat. Nos. 6,924,266; 7,241,738;7,317,077; 7,408,021; 7,745,572; 8,067,378; 8,293,703; 8,569,446; and8,716,247, and U.S. Patent Application Publication Nos. 2017/0360885;2017/0020957; 2016/0361380; and 2016/0215031. The disclosures of thesepatents and published applications are incorporated by reference hereinin their entirety.

FT is represented by:

SEQ ID NO. 1: IDQQVLSRIKLEIKRCL orIle-Asp-Gln-Gln-Val-Leu-Ser-Arg-Ile-Lys-Leu-Glu- Ile-Lys-Arg-Cys-Leu.

The term “fragment” refers to a protein or polypeptide that consists ofa continuous subsequence of the amino acid sequence of a protein orpeptide and includes naturally occurring fragments such as splicevariants and fragments resulting from naturally occurring in vivoprotease activity. Such a fragment may be truncated at the aminoterminus, the carboxy terminus, and/or internally (such as by naturalsplicing). Such fragments may be prepared with or without an aminoterminal methionine. The term “fragment” includes fragments, whetheridentical or different, from the same protein or peptide, with acontiguous amino acid sequence in common or not, joined together, eitherdirectly or through a linker. A person having ordinary skill in the artwill be capable of selecting a suitable fragment for use in theembodiments without undue experimentation using the guidelines andprocedures outlined herein.

The term “variant” refers to a protein or polypeptide in which one ormore amino acid substitutions, deletions, and/or insertions are presentas compared to the amino acid sequence of an protein or peptide andincludes naturally occurring allelic variants or alternative splicevariants of an protein or peptide. The term “variant” includes thereplacement of one or more amino acids in a peptide sequence with asimilar or homologous amino acid(s) or a dissimilar amino acid(s). Thereare many scales on which amino acids can be ranked as similar orhomologous. (Gunnar von Heijne, Sequence Analysis in Molecular Biology,p. 123-39 (Academic Press, New York, N.Y. 1987.) Preferred variantsinclude alanine substitutions at one or more of amino acid positions.Other preferred substitutions include conservative substitutions thathave little or no effect on the overall net charge, polarity, orhydrophobicity of the protein. Conservative substitutions are set forthin Table 2 below.

TABLE 2 Conservative Amino Acid Substitutions Basic: arginine lysinehistidine Acidic: glutamic acid aspartic acid Uncharged Polar: glutamineasparagine serine threonine tyrosine Non-Polar: phenylalanine tryptophancysteine glycine alanine valine praline methionine leucine isoleucineTable 3 sets out another scheme of amino acid substitution:

TABLE 3 Original Residue Substitutions Ala gly; ser Arg lys Asn gln; hisAsp glu Cys ser Gln asn Glu asp Gly ala; pro His asn; gln Ile eu; valLeu ile; val Lys arg; gln; glu Met leu; tyr; ile Phe met; leu; tyr Serthr Thr ser Trp tyr Tyr trp; phe Val ile; leu

Other variants can consist of less conservative amino acidsubstitutions, such as selecting residues that differ more significantlyin their effect on maintaining (a) the structure of the polypeptidebackbone in the area of the substitution, for example, as a sheet orhelical conformation, (b) the charge or hydrophobicity of the moleculeat the target site, or (c) the bulk of the side chain. The substitutionsthat in general are expected to have a more significant effect onfunction are those in which (a) glycine and/or proline is substituted byanother amino acid or is deleted or inserted; (b) a hydrophilic residue,e.g., seryl or threonyl, is substituted for (or by) a hydrophobicresidue, e.g., leucyl, isoleucyl, phenylalanyl, valyl, or alanyl; (c) acysteine residue is substituted for (or by) any other residue; (d) aresidue having an electropositive side chain, e.g., lysyl, arginyl, orhistidyl, is substituted for (or by) a residue having an electronegativecharge, e.g., glutamyl or aspartyl; or (e) a residue having a bulky sidechain, e.g., phenylalanine, is substituted for (or by) one not havingsuch a side chain, e.g., glycine. Other variants include those designedto either generate a novel glycosylation and/or phosphorylation site(s),or those designed to delete an existing glycosylation and/orphosphorylation site(s). Variants include at least one amino acidsubstitution at a glycosylation site, a proteolytic cleavage site and/ora cysteine residue. Variants also include proteins and peptides withadditional amino acid residues before or after the protein or peptideamino acid sequence on linker peptides. For example, a cysteine residuemay be added at both the amino and carboxy terminals of a peptide inorder to allow the cyclisation of the peptide by the formation of adi-sulphide bond. The term “variant” also encompasses polypeptides thathave the amino acid sequence of a peptide with at least one and up to 25or more additional amino acids flanking either the 3′ or 5′ end of thepeptide.

The term “derivative” refers to a chemically modified protein orpolypeptide that has been chemically modified either by naturalprocesses, such as processing and other post-translationalmodifications, but also by chemical modification techniques, as forexample, by addition of one or more polyethylene glycol molecules,sugars, phosphates, and/or other such molecules, where the molecule ormolecules are not naturally attached to wild-type proteins or peptides.Derivatives include salts. Such chemical modifications are welldescribed in basic texts and in more detailed monographs, as well as ina voluminous research literature, and they are well known to those ofskill in the art. It will be appreciated that the same type ofmodification may be present in the same or varying degree at severalsites in a given protein or polypeptide. Also, a given protein orpolypeptide may contain many types of modifications. Modifications canoccur anywhere in a protein or polypeptide, including the peptidebackbone, the amino acid side-chains, and the amino or carboxyl termini.Modifications include, for example, acetylation, acylation,ADP-ribosylation, amidation, covalent attachment of flavin, covalentattachment of a heme moiety, covalent attachment of a nucleotide ornucleotide derivative, covalent attachment of a lipid or lipidderivative, covalent attachment of phosphotidylinositol, cross-linking,cyclization, disulfide bond formation, demethylation, formation ofcovalent cross-links, formation of cysteine, formation of pyroglutamate,formylation, gamma-carboxylation, glycosylation, GPI anchor formation,hydroxylation, iodination, methylation, myristoylation, oxidation,proteolytic processing, phosphorylation, prenylation, racemization,glycosylation, lipid attachment, sulfation, gamma-carboxylation ofglutamic acid residues, hydroxylation and ADP-ribosylation,selenoylation, sulfation, transfer-RNA mediated addition of amino acidsto proteins, such as arginylation, and ubiquitination. See, forinstance, Proteins—Structure And Molecular Properties, 2nd Ed., T. E.Creighton, W. H. Freeman and Company, New York (1993) and Wold, F.,“Posttranslational Protein Modifications: Perspectives and Prospects,”pgs. 1-12 in Posttranslational Covalent Modification Of Proteins, B. C.Johnson, Ed., Academic Press, New York (1983); Seifter et al., Meth.Enzymol. 182:626-646 (1990) and Rattan et al., “Protein Synthesis:Posttranslational Modifications and Aging,” Ann. N.Y. Acad. Sci. 663:48-62 (1992). The term “derivatives” include chemical modificationsresulting in the protein or polypeptide becoming branched or cyclic,with or without branching. Cyclic, branched and branched circularproteins or polypeptides may result from post-translational naturalprocesses and may be made by entirely synthetic methods, as well.

The term “homologue” refers to a protein that is at least 60 percentidentical in its amino acid sequence of peptide as determined bystandard methods that are commonly used to compare the similarity inposition of the amino acids of two polypeptides. The degree ofsimilarity or identity between two proteins can be readily calculated byknown methods, including but not limited to those described inComputational Molecular Biology, Lesk, A. M., ed., Oxford UniversityPress, New York, 1988; Biocomputing: Informatics and Genome Projects,Smith, D. W., ed., Academic Press, New York, 1993; Computer Analysis ofSequence Data, Part I, Griffin, A. M., and Griffin, H. G., eds., HumanaPress, New Jersey, 1994; Sequence Analysis in Molecular Biology, vonHeinje, G., Academic Press, 1987; Sequence Analysis Primer, Gribskov, M.and Devereux, J., eds., M Stockton Press, New York, 1991; and Carillo H.and Lipman, D., SIAM, J. Applied Math., 48:1073 (1988). Preferredmethods to determine identity are designed to give the largest matchbetween the sequences tested. Methods to determine identity andsimilarity are codified in publicly available computer programs.

Preferred computer program methods useful in determining the identityand similarity between two sequences include, but are not limited to,the GCG program package (Devereux, J., et al., Nucleic Acids Research,12(1): 387 (1984)), BLASTP, BLASTN, and FASTA, Atschul, S. F. et al., J.Molec. Biol., 215: 403-410 (1990). The BLAST X program is publiclyavailable from NCBI and other sources (BLAST Manual, Altschul, S., etal., NCBI NLM NIH Bethesda, Md. 20894; Altschul, S., et al., J. Mol.Biol., 215: 403-410 (1990). By way of example, using a computeralgorithm such as GAP (Genetic Computer Group, University of Wisconsin,Madison, Wis.), the two proteins or polypeptides for which the percentsequence identity is to be determined are aligned for optimal matchingof their respective amino acids (the “matched span”, as determined bythe algorithm).

A gap opening penalty (which is calculated as 3 times the averagediagonal; the “average diagonal” is the average of the diagonal of thecomparison matrix being used; the “diagonal” is the score or numberassigned to each perfect amino acid match by the particular comparisonmatrix) and a gap extension penalty (which is usually 1/10 times the gapopening penalty), as well as a comparison matrix such as PAM 250 orBLOSUM 62 are used in conjunction with the algorithm. A standardcomparison matrix (see Dayhoff et al. in: Atlas of Protein Sequence andStructure, vol. 5, supp.3 for the PAM250 comparison matrix; see Henikoffet al., Proc. Natl. Acad. Sci USA, 89:10915-10919 for the BLOSUM 62comparison matrix) also may be used by the algorithm. The percentidentity then is calculated by the algorithm. Homologues will typicallyhave one or more amino acid substitutions, deletions, and/or insertionsas compared with the comparison protein or peptide, as the case may be.

The term “fusion protein” refers to a protein where one or more peptidesare recombinantly fused or chemically conjugated (including covalentlyand non-covalently) to a protein such as (but not limited to) anantibody or antibody fragment like an Fab fragment or short chain Fv.The term “fusion protein” also refers to multimers (i.e. dimers,trimers, tetramers and higher multimers) of peptides. Such multimerscomprise homomeric multimers comprising one peptide, heteromericmultimers comprising more than one peptide, and heteromeric multimerscomprising at least one peptide and at least one other protein. Suchmultimers may be the result of hydrophobic, hyrdrophilic, ionic and/orcovalent associations, bonds or links, may be formed by cross-linksusing linker molecules or may be linked indirectly by, for example,liposome formation

The term “peptide mimetic” or “mimetic” refers to biologically activecompounds that mimic the biological activity of a peptide or a proteinbut are no longer peptidic in chemical nature, that is, they no longercontain any peptide bonds (that is, amide bonds between amino acids).Here, the term peptide mimetic is used in a broader sense to includemolecules that are no longer completely peptidic in nature, such aspseudo-peptides, semi-peptides and peptoids. Examples of peptidemimetics in this broader sense (where part of a peptide is replaced by astructure lacking peptide bonds) are described below. Whether completelyor partially non-peptide, peptide mimetics according to the embodimentsprovide a spatial arrangement of reactive chemical moieties that closelyresemble the three-dimensional arrangement of active groups in thepeptide on which the peptide mimetic is based. As a result of thissimilar active-site geometry, the peptide mimetic has effects onbiological systems that are similar to the biological activity of thepeptide.

The peptide mimetics of the embodiments are preferably substantiallysimilar in both three-dimensional shape and biological activity to thepeptides described herein. Examples of methods of structurally modifyinga peptide known in the art to create a peptide mimetic include theinversion of backbone chiral centers leading to D-amino acid residuestructures that may, particularly at the N-terminus, lead to enhancedstability for proteolytical degradation without adversely affectingactivity. An example is given in the paper “Tritriated D-ala₁-Peptide TBinding”, Smith C. S. et al., Drug Development Res., 15, pp. 371-379(1988). A second method is altering cyclic structure for stability, suchas N to C interchain imides and lactames (Ede et al. in Smith and Rivier(Eds.) “Peptides: Chemistry and Biology”, Escom, Leiden (1991), pp.268-270). An example of this is given in conformationally restrictedthymopentin-like compounds, such as those disclosed in U.S. Pat. No.4,457,489 (1985), Goldstein, G. et al., the disclosure of which isincorporated by reference herein in its entirety. A third method is tosubstitute peptide bonds in the peptide by pseudopeptide bonds that.confer resistance to proteolysis.

A number of pseudopeptide bonds have been described that in general donot affect peptide structure and biological activity. One example ofthis approach is to substitute retro-inverso pseudopeptide bonds(“Biologically active retroinverso analogues of thymopentin”, Sisto A.et al in Rivier, J. E. and Marshall, G. R. (eds) “Peptides, Chemistry,Structure and Biology”, Escom, Leiden (1990), pp. 722-773) and Dalpozzo,et al. (1993), Int. J. Peptide Protein Res., 41:561-566, incorporatedherein by reference). According to this modification, the amino acidsequences of the peptides may be identical to the sequences of a peptidedescribed above, except that one or more of the peptide bonds arereplaced by a retro-inverso pseudopeptide bond. Preferably the mostN-terminal peptide bond is substituted, since such a substitution willconfer resistance to proteolysis by exopeptidases acting on theN-terminus. Further modifications also can be made by replacing chemicalgroups of the amino acids with other chemical groups of similarstructure. Another suitable pseudopeptide bond that is known to enhancestability to enzymatic cleavage with no or little loss of biologicalactivity is the reduced isostere pseudopeptide bond (Couder, et al.(1993), Int. J. Peptide Protein Res., 41:181-184, incorporated herein byreference in its entirety).

Thus, the amino acid sequences of these peptides may be identical to thesequences of a peptide, except that one or more of the peptide bonds arereplaced by an isostere pseudopeptide bond. Preferably the mostN-terminal peptide bond is substituted, since such a substitution wouldconfer resistance to proteolysis by exopeptidases acting on theN-terminus. The synthesis of peptides with one or more reduced isosterepseudopeptide bonds is known in the art (Couder, et al. (1993), citedabove). Other examples include the introduction of ketomethylene ormethylsulfide bonds to replace peptide bonds.

Peptoid derivatives of peptides represent another class of peptidemimetics that retain the important structural determinants forbiological activity, yet eliminate the peptide bonds, thereby conferringresistance to proteolysis (Simon, et al., 1992, Proc. Natl. Acad. Sci.USA, 89:9367-9371, incorporated herein by reference in its entirety).Peptoids are oligomers of N-substituted glycines. A number of N-alkylgroups have been described, each corresponding to the side chain of anatural amino acid (Simon, et al. (1992), cited above). Some or all ofthe amino acids of the peptides may be replaced with the N-substitutedglycine corresponding to the replaced amino acid.

The term “peptide mimetic” or “mimetic” also includes reverse-D peptidesand enantiomers as defined below.

The term “reverse-D peptide” refers to a biologically active protein orpeptide consisting of D-amino acids arranged in a reverse order ascompared to the L-amino acid sequence of an peptide. Thus, the carboxyterminal residue of an L-amino acid peptide becomes the amino terminalfor the D-amino acid peptide and so forth. For example, the peptide,ETESH, becomes H_(d)S_(d)E_(d)T_(d)E_(d), where E_(d), H_(d), S_(d), andT_(d) are the D-amino acids corresponding to the L-amino acids, E, H, S,and T respectively.

The term “enantiomer” refers to a biologically active protein or peptidewhere one or more the L-amino acid residues in the amino acid sequenceof a peptide is replaced with the corresponding D-amino acid residue(s).

A “composition” as used herein, refers broadly to any compositioncontaining a recited peptide or amino acid sequence and, optionally anadditional active agent. The composition may comprise a dry formulation,an aqueous solution, or a sterile composition. Compositions comprisingpeptides may be employed as hybridization probes. The probes may bestored in freeze-dried form and may be associated with a stabilizingagent such as a carbohydrate. In hybridizations, the probe may bedeployed in an aqueous solution containing salts, e.g., NaCl,detergents, e.g. sodium dodecyl sulfate (SDS), and other components,e.g., Denhardt's solution, dry milk, salmon sperm DNA, etc.

The expression “low grade prostate cancer” denotes prostate cancerpresenting a biopsy of prostate tissue, i.e., a single foci or multiplefoci, having the highest Gleason grade of ≤6 or 3+3 that was detected bybiopsy. It will be understood that biopsies procedures, which often takenumerous samples from the prostate gland, do not sample the entire glandand consequently, there may be other foci present that were notdetected. The expression “low grade unifocal prostate cancer” denotes asingle cancerous focus having a Gleason grade of ≤6, or 3+3. Theexpression “progression of prostate cancer” typically denotes a higherGleason grade in any single focus (the highest grade in all biopsiestaken together is the grade), but also denotes greater amounts of cancerin the biopsy (i.e. a higher percentage of cancer in a given biopsyfocus if it gets past 50% for example; or more foci positive forcancer). For example, if a patient proceeds from at one point in timewith one positive core sample having 5% tumor, to at a later point intime, having 5 cores with each having 40% tumor (all of same Gleasongrade), this would be considered progression, although it would not be“Gleason grade progression.” On the other hand, “Gleason gradeprogression” would be present if a patient proceeded from 4 cores ofGrade 6 each with 40% tumor, and then subsequently had only one positivecore with 5% tumor, but Gleason grade 7, then that progression would beconsidered “Gleason grade progression.”

When referring to a “biopsy”, those skilled in the art will appreciatethat a typical biopsy consists of multiple “quadrant” samples, generallyat least 10 or 12, sampling all areas of the gland (left and right;apex, mid, and base for each; and medial and lateral for each, andtransition L and R), thus equaling 14 zones. Therefore a reference to“biopsy” or “a biopsy” denotes 10-15 biopsies at the same time, witheach being reported separately.

Throughout this description, the expression “multifocal cancer” denotesone or more cancers selected from prostate cancer, squamous cell headand neck carcinoma (HNSCC), cutaneous squamous cell carcinoma (skinSCC), breast cancer, adenocarcinoma and SCC of lung, esopahgeal cancer,gastric cancer, colon cancer, bladder cancer, cervical cancer, melanoma,brain cancer, pancreatic cancer, ovarian cancer, bone marrow cancer, andleukemia. Throughout this description, the term “prevention,” orvariants thereof, do not necessarily mean complete prevention in allcases, but instead refers to preventing the development or incidence ofmultifocal cancer, when compared to controls such as activesurveillance. For example, assume that 60% of a group of patients havingunifocal cancer, or a low grade, low risk, cancer, developed multifocalcancer with no treatment (or treatment with a control) over a period of36 months, but only 20% developed multifocal cancer over a period of 36months when treated in accordance with the embodiments. The treatmentsdescribed herein therefore would be effective in preventing a largenumber of patients from developing multifocal cancer, who otherwisewould have developed multifocal cancer if not treated in accordance withthe embodiments.

In an embodiment in which an additional active agent is used togetherwith composition, the expression “active agent” is used to denote anyagent capable of removing unwanted cellular proliferations and/or tissuegrowth. Suitable active agents may include, but are not limited to: (i)anti-cancer active agents (such as alkylating agents, topoisomerase Iinhibitors, topoisomerase II inhibitors, RNA/DNA antimetabolites, andantimitotic agents); (ii) active agents for treating benign growths suchas anti-acne and anti-wart active agents; (iii) antiandrogen compounds,(cyproterone acetate(1α,2ß-methylene-6-chloro-17α-acetoxy-6-dehydroprogesterone) Tamoxifen,aromatase inhibitors); (iv) alpha1-adrenergic receptor blockers(tamsulosin, terazosin, doxazosin, prazosin, bunazosin, indoramin,alfulzosin, silodosin); (v) 5α-reductase inhibitors (finasteride,dutasteride); (vi) phosphodiesterase type 5 (PDE5) inhibitors(tadalafil) and combinations thereof.

The embodiments are directed to methods of administering to a low grade,low risk, localized prostate cancer tumor a composition comprising atleast one pharmaceutically active ingredient capable of inducingnecrosis of the low grade, low risk, localized prostate cancer tumor,and reducing prostate cancer incidence, reducing prostate cancer grade,and reducing prostate cancer progression (worsening) in the entirehemi-prostate where the initial unifocal tumor was located and treated.Suitable pharmaceutically active ingredients capable of inducingnecrosis of such tumors, include, for example, fexapotide triflutate,(FT), a peptide described by the amino acid sequenceIle-Asp-Gln-Gln-Val-Leu-Ser-Arg-Ile-Lys-Leu-Glu-Ile-Lys-Arg-Cys-Leu (SEQID NO. 1), ZYTIGA® (abiraterone acetate), EARLEADA® (apalutamide),abazitaxel, CASODEX® (Bicalutamide), ELIGARD® and LUPRON®, (LeuprolideAcetate), FIRMAGON® (Degarelix), flutamide, JEVTANA® (Cabazitaxel),mitoxantrone hydrochloride, NILANDRON® (Nilutamide), PROVENGE®(Sipuleucel-T), TAXOTERE® (Docetaxel), XOFIGO® (Radium 223 Dichloride),XTANDI® (Enzalutamide), ZOLADEX® (Goserelin Acetate), and mixtures andcombinations thereof. In an embodiment, the compositions areadministered more than once. The embodiments therefore provide a methodof reducing prostate cancer incidence, grade, and progression in aminimally invasive manner, by administering the compositions to mammalsthat typically would not be treated. It is generally accepted thatmammals with low grade unifocal or multifocal prostate cancer havingGleason grade 6 undergo active surveillance (AS), or no treatment. See,e.g., Ahmed, et al., “Do Low-Grade and Low-Volume Prostate Cancers Bearthe Hallmarks of Malignancy,” thelancet.com, pp e509-e517 (2012).

The inventor unexpectedly discovered that administration of suchcompositions to a single focus of a multifocal cancer significantlyreduced multifocal cancer incidence, cancer grade, and cancerprogression in the entire organ or organism. The methods of theembodiments therefore provide a non-invasive method of reducingmultifocal cancer, when compared to organ removal or transplantation, oreven focal ablation, excision, chemotherapy, or radiation. Even activesurveillance requires multiple and repeated prostate biopsies andevaluation, and places a large burden on the health care system.Accordingly, the methods described herein are useful in retarding theincidence, occurrence, and progression of multifocal cancer in anon-invasive manner.

With respect to prostate cancer, and in contrast to the publishedliterature, mammals treated with the compositions of the presentinvention exhibited a dramatic decrease in the incidence of multifocalprostate cancer, a dramatic decrease in multifocal Gleason gradeincrease, and a dramatic decrease in multifocal prostate cancerprogression in the entire prostate. The method of the embodiments canreduce entire prostate multifocal cancer with Gleason upgrade (patientswith >=1 new focus in entire prostate with increased Gleason grade),when compared to active surveillance controls, when measured at least 18months after treatment, by an amount of from about 40% to about 100%, orfrom about 50% to about 90%, or from about 50% to about 80%, or anyvalue therebetween. The method of the embodiments can reduce entireprostate multifocal cancer with Gleason upgrade, when compared to activesurveillance controls, when measured at least 36 months after treatment,by an amount of from about 30% to about 90%, or from about 45% to about90%, or from about 47% to about 80%, or any value therebetween. Themethod of the embodiments can reduce entire prostate multifocal cancerwith Gleason upgrade, when compared to active surveillance controls,when measured at least 48 months after treatment, by an amount of fromabout 5% to about 85%, or from about 10% to about 70%, or from about 12%to about 65%, or any value therebetween.

The method of the embodiments can reduce entire prostate multifocalcancer with Gleason grade primary pattern ≥4 (patients with >=1 newfocus in entire prostate with an increase in Gleason grade primarypattern), when compared to active surveillance controls, when measuredat least 18 months after treatment, by an amount of from about 40% toabout 100%, or from about 70% to about 100%, or from about 80% to about100%, or any value therebetween. The method of the embodiments canreduce entire prostate multifocal cancer with Gleason grade primarypattern ≥4, when compared to active surveillance controls, when measuredat least 36 months after treatment, by an amount of from about 30% toabout 100%, or from about 70% to about 100%, or from about 80% to about100%, or any value therebetween. The method of the embodiments canreduce entire prostate multifocal cancer with Gleason grade primarypattern ≥4, when compared to active surveillance controls, when measuredat least 48 months after treatment, by an amount of from 20% to about100%, or from about 50% to about 100%, or from about 60% to about 100%,or any value therebetween.

The method of the embodiments can reduce multifocal prostate cancer inthe lobe (or hemi-prostate) in which the foci were initially treatedwith Gleason upgrade (patients with >=1 new focus in treated lobe orhemi-prostate with increased Gleason grade), when compared to activesurveillance controls, when measured at least 18 months after treatment,by an amount of from about 40% to about 100%, or from about 50% to about90%, or from about 60% to about 80%, or any value therebetween. Themethod of the embodiments can reduce hemi-prostate multifocal cancerwith Gleason upgrade, when compared to active surveillance controls,when measured at least 36 months after treatment, by an amount of fromabout 50% to about 90%, or from about 50% to about 85%, or from about50% to about 80%, or any value therebetween. The method of theembodiments can reduce hemi-prostate multifocal cancer with Gleasonupgrade, when compared to active surveillance controls, when measured atleast 48 months after treatment, by an amount of from about 15% to about80%, or from about 15% to about 75%, or from about 16% to about 72%, orany value therebetween.

The method of the embodiments can reduce multifocal prostate cancer inthe lobe (or hemi-prostate) in which the foci were initially treatedwith Gleason primary pattern ≥4 (patients with >=1 new focus in treatedlobe or hemi-prostate with an increase in Gleason grade primarypattern), when compared to active surveillance controls, when measuredat least 18 months after treatment, by an amount of from about 50% toabout 100%, or from about 70% to about 100%, or from about 75% to about100%, or any value therebetween. The method of the embodiments canreduce hemi-prostate multifocal cancer with Gleason primary pattern ≥4,when compared to active surveillance controls, when measured at least 36months after treatment, by an amount of from about 45% to about 100%, orfrom about 70% to about 100%, or from about 75% to about 100%, or anyvalue therebetween. The method of the embodiments can reducehemi-prostate multifocal cancer with Gleason upgrade (patients with >=1new focus in entire prostate with increased Gleason grade), whencompared to active surveillance controls, when measured at least 48months after treatment, by an amount of from 45% to about 100%, or fromabout 60% to about 100%, or from about 70% to about 100%, or any valuetherebetween.

The method of the embodiments can reduce the percentage of mammalshaving conventional cancer treatment (surgery, radiotherapy, orchemotherapy), and exhibiting ≥1 new focus in the entire prostate withan increase in Gleason grade (cancer treatment with new multifocal withGleason upgrade), when compared to active surveillance controls, whenmeasured at least 18 months after treatment, by an amount of from about50% to about 100%, or from about 55% to about 90%, or from about 60% toabout 80%, or any value therebetween. The method of the embodiments canreduce cancer treatment with new multifocal with Gleason upgrade in theentire prostate, when compared to active surveillance controls, whenmeasured at least 36 months after treatment, by an amount of from about40% to about 90%, or from about 45% to about 90%, or from about 50% toabout 80%, or any value therebetween. The method of the embodiments canreduce cancer treatment with new multifocal with Gleason upgrade in theentire prostate, when compared to active surveillance controls, whenmeasured at least 48 months after treatment, by an amount of from about35% to about 85%, or from about 40% to about 85%, or from about 48% toabout 84%, or any value therebetween.

The method of the embodiments can reduce the percentage of mammalshaving conventional cancer treatment (surgery, radiotherapy, orchemotherapy), and exhibiting entire prostate multifocal cancer withGleason grade primary pattern ≥4 (cancer treatment with new multifocaland an increase in Gleason grade primary pattern), when compared toactive surveillance controls, when measured at least 18 months aftertreatment, by an amount of from about 70% to about 100%, or from about70% to about 100%, or from about 80% to about 100%, or any valuetherebetween. The method of the embodiments can reduce cancer treatmentwith new multifocal and an increase in Gleason grade primary pattern,when compared to active surveillance controls, when measured at least 36months after treatment, by an amount of from about 60% to about 100%, orfrom about 70% to about 100%, or from about 80% to about 100%, or anyvalue therebetween. The method of the embodiments can reduce cancertreatment with new multifocal and an increase in Gleason grade primarypattern, when compared to active surveillance controls, when measured atleast 48 months after treatment, by an amount of from 50% to about 100%,or from about 70% to about 100%, or from about 80% to about 100%, or anyvalue therebetween.

The method of the embodiments can reduce the percentage of mammalshaving conventional cancer treatment (surgery, radiotherapy, orchemotherapy), and having multifocal prostate cancer in the lobe (orhemi-prostate) in which the foci were initially treated with Gleasonupgrade (cancer treatment with new multifocal with increased Gleasongrade in the hemi-prostate), when compared to active surveillancecontrols, when measured at least 18 months after treatment, by an amountof from about 40% to about 100%, or from about 50% to about 90%, or fromabout 60% to about 75%, or any value therebetween. The method of theembodiments can reduce cancer treatment with new multifocal withincreased Gleason grade in the hemi-prostate, when compared to activesurveillance controls, when measured at least 36 months after treatment,by an amount of from about 50% to about 90%, or from about 55% to about75%, or from about 50% to about 80%, or any value therebetween. Themethod of the embodiments can reduce cancer treatment with newmultifocal with increased Gleason grade in the hemi-prostate, whencompared to active surveillance controls, when measured at least 48months after treatment, by an amount of from about 15% to about 80%, orfrom about 35% to about 75%, or from about 40% to about 75%, or anyvalue therebetween.

The method of the embodiments can reduce the percentage of mammalshaving conventional cancer treatment (surgery, radiotherapy, orchemotherapy), and having multifocal prostate cancer in the lobe (orhemi-prostate) in which the foci were initially treated with Gleasonprimary pattern ≥4 (cancer treatment with new multifocal and an increasein Gleason grade primary pattern in the hemi-prostate), when compared toactive surveillance controls, when measured at least 18 months aftertreatment, by an amount of from about 65% to about 100%, or from about70% to about 100%, or from about 75% to about 100%, or any valuetherebetween. The method of the embodiments can reduce cancer treatmentwith new multifocal and an increase in Gleason grade primary pattern inthe hemi-prostate, when compared to active surveillance controls, whenmeasured at least 36 months after treatment, by an amount of from about65% to about 100%, or from about 70% to about 100%, or from about 75% toabout 100%, or any value therebetween. The method of the embodiments canreduce cancer treatment with new multifocal and an increase in Gleasongrade primary pattern in the hemi-prostate, when compared to activesurveillance controls, when measured at least 48 months after treatment,by an amount of from 60% to about 100%, or from about 65% to about 100%,or from about 75% to about 100%, or any value therebetween.

The embodiments include a method of treating a mammal having low gradeunifocal prostate cancer, comprising administering once or more thanonce a composition comprising FT to a single low grade, low-riskprostate cancer focus present in the mammal's prostate, either alone orin combination with administration of an additional active agent. Themethod includes, but is not limited to, administering the compositionintramuscularly, orally, intravenously, intraperitoneally,intracerebrally (intraparenchymally), intracerebroyentricularly,intralesionally, intraocularly, intraarterially, intrathecally,intratumorally, intranasally, topically, transdermally, subcutaneously,or intradermally, either alone or conjugated to a carrier.

Any mammal can benefit from use of the invention, including humans,mice, rabbits, dogs, sheep and other livestock, any mammal treated ortreatable by a veterinarian, zoo-keeper, or wildlife preserve employee.Preferred mammals are humans, sheep, and dogs. Throughout thisdescription mammals and patients are used interchangeably.

It will be apparent to one of skill in the art that other smallerfragments of FT may be selected such that these peptides will possessthe same or similar biological activity. Other fragments of FT may beselected by one skilled in the art such that these peptides will possessthe same or similar biological activity. The term “FT” as used in theembodiments therefore encompasses these other fragments. In general, thepeptides of the embodiments have at least 4 amino acids, preferably atleast 5 amino acids, and more preferably at least 6 amino acids.

The embodiments also encompass methods of treatment comprisingadministering a composition comprising FT that includes two or more FTsequences joined together, together with an additional active agent. Tothe extent that FT has the desired biological activity, it follows thattwo or more FT sequences would also possess the desired biologicalactivity.

FT and fragments, variants, derivatives, homologues, fusion proteins andmimetics thereof encompassed by this embodiment can be prepared usingmethods known to those of skill in the art, such as recombinant DNAtechnology, protein synthesis and isolation of naturally occurringpeptides, proteins, variants, derivatives and homologues thereof. FT andfragments, variants, derivatives, homologues, fusion proteins andmimetics thereof can be prepared from other peptides, proteins, andfragments, variants, derivatives and homologues thereof using methodsknown to those having skill in the art. Such methods include (but arenot limited to) the use of proteases to cleave the peptide, or proteininto FT. Any method disclosed in, for example, U.S. Pat. Nos. 6,924,266;7,241,738; 7,317,077; 7,408,021; 7,745,572; 8,067,378; 8,293,703;8,569,446; and 8,716,247, and U.S. Patent Application Publication Nos.2017/0360885; 2017/0020957; 2016/0361380; and 2016/0215031, can be usedto prepare the FT peptide described herein. The disclosures of thesepatent documents are incorporated by reference herein in theirentireties.

The additional active agent, if used in addition to the one or morepharmaceutically active ingredients, can be one or more active agentsselected from (i) anti-cancer active agents (such as alkylating agents,topoisomerase I inhibitors, topoisomerase II inhibitors, RNA/DNAantimetabolites, and antimitotic agents); (ii) active agents fortreating benign growths such as anti-acne and anti-wart active agents(salicylic acid); (iii) antiandrogen compounds, (cyproterone acetate(1α,2β-methylene-6-chloro-17α-acetoxy-6-dehydroprogesterone)) Tamoxifen,aromatase inhibitors); (iv) alpha1-adrenergic receptor blockers(tamsulosin, terazosin, doxazosin, prazosin, bunazosin, indoramin,alfulzosin, silodosin); (v) 5α-reductase inhibitors (finasteride,dutasteride); (vi) phosphodiesterase type 5 (PDE5) inhibitors(tadalafil) and combinations thereof. Preferably, the additional agentis an anti-cancer agent and specifically an agent useful in treatingprostate cancer.

Therapeutic compositions described herein may comprise an amount ofpharmaceutically active ingredient in admixture with a pharmaceuticallyacceptable carrier. In some alternative embodiments, the additionalactive agent can be administered in the same composition with thepharmaceutically active ingredient, and in other embodiments, thecomposition comprising the pharmaceutically active ingredient isadministered as an injection, whereas the additional active agent isformulated into an oral medication (gel, capsule, tablet, liquid, etc.).The carrier material may be water for injection, preferably supplementedwith other materials common in solutions for administration to mammals.Typically, when the pharmaceutically active ingredient is FT, it will beadministered in the form of a composition comprising the purified FTpeptide (or chemically synthesized FT peptide) in conjunction with oneor more physiologically acceptable carriers, excipients, or diluents.Neutral buffered saline or saline mixed with serum albumin are exemplaryappropriate carriers. Preferably, the product is formulated as alyophilizate using appropriate excipients (e.g., sucrose). Otherstandard carriers, diluents, and excipients may be included as desired.Compositions of the embodiments also may comprise buffers known to thosehaving ordinary skill in the art with an appropriate range of pH values,including Tris buffer of about pH 7.0-8.5, or acetate buffer of about pH4.0-5.5, which may further include sorbitol or a suitable substitutetherefor.

Solid dosage forms for oral administration include but are not limitedto, capsules, tablets, pills, powders, and granules. In such soliddosage forms, the additional active agent, and/or the pharmaceuticallyactive ingredient can be admixed with at least one of the following: (a)one or more inert excipients (or carrier), such as sodium citrate ordicalcium phosphate; (b) fillers or extenders, such as starches,lactose, sucrose, glucose, mannitol, and silicic acid; (c) binders, suchas carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone,sucrose and acacia; (d) humectants, such as glycerol; (e) disintegratingagents, such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain complex silicates, and sodium carbonate; (f)solution retarders, such as paraffin; (g) absorption accelerators, suchas quaternary ammonium compounds; (h) wetting agents, such as acetylalcohol and glycerol monostearate; (i) adsorbents, such as kaolin andbentonite; and (j) lubricants, such as talc, calcium stearate, magnesiumstearate, solid polyethylene glycols, sodium lauryl sulfate, or mixturesthereof. For capsules, tablets, and pills, the dosage forms may alsocomprise buffering agents.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, solutions, suspensions, syrups, and elixirs. Inaddition to the active compounds, the liquid dosage forms may compriseinert diluents commonly used in the art, such as water or othersolvents, solubilizing agents, and emulsifiers. Exemplary emulsifiersare ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate,benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butyleneglycol,dimethylformamide, oils, such as cottonseed oil, groundnut oil, corngerm oil, olive oil, castor oil, and sesame oil, glycerol,tetrahydrofurfuryl alcohol, polyethyleneglycols, fatty acid esters ofsorbitan, or mixtures of these substances, and the like.

Besides such inert diluents, the composition can also include adjuvants,such as wetting agents, emulsifying and suspending agents, sweetening,flavoring, and perfuming agents.

Actual dosage levels of active ingredients in the compositions of theembodiments may be varied to obtain an amount of the pharmaceuticallyactive ingredient and additional active agent that is effective toobtain a desired therapeutic response for a particular composition. Theselected dosage level therefore depends upon the desired therapeuticeffect, the route of administration, the desired duration of treatment,and other factors.

With mammals, including humans, the effective amounts can beadministered on the basis of body surface area. The interrelationship ofdosages for animals of various sizes, species and humans (based on mg/M²of body surface) is described by E. J. Freireich et al., CancerChemother. Rep., 50 (4):219 (1966). Body surface area may beapproximately determined from the height and weight of an individual(see e.g., Scientific Tables, Geigy Pharmaceuticals, Ardsley, N.Y. pp.537-538 (1970)).

It will be understood that the specific dose level for any particularpatient will depend upon a variety of factors including the body weight,general health, sex, diet, time and route of administration, potency ofthe administered drug, rates of absorption and excretion, combinationwith other drugs and the severity of the particular disease beingtreated.

A method of administering a composition comprising the pharmaceuticallyactive ingredient according to the embodiments includes, but is notlimited to, administering the compositions intramuscularly, orally,intravenously, intraperitoneally, intracerebrally (intraparenchymally),intracerebroventricularly, intratumorally, intralesionally,intradermally, intrathecally, intranasally, intraocularly,intraarterially, topically, transrectally, transperitoneally,transdermally, via an aerosol, infusion, bolus injection, implantationdevice, sustained release system etc. Any method of administrationdisclosed in, for example, U.S. Pat. Nos. 6,924,266; 7,241,738;7,317,077; 7,408,021; 7,745,572; 8,067,378; 8,293,703; 8,569,446; and8,716,247, and U.S. Patent Application Publication Nos. 2017/0360885;2017/0020957; 2016/0361380; and 2016/0215031, can be used.

Use of FT is a preferred embodiment. FT is a new molecular entity whichin vitro stimulates caspase pathways (activation of caspases 7, 8, and10, caspase recruitment domains 6, 11, and 14, and DIABLO), tumornecrosis factor pathways (activation of TNF1, TNFSF6, TNFSF8, TNFSF9,CD70 ligands, and TNFRSF19L, TNFRSF25, TRAF2, TRAF3, TRAF4, TRAF6receptors), and BCL pathways (activation of BIK, HRK, BCL2L10 and BCL3)in prostate glandular epithelial cells, based on tissue culture geneticarray data. FT selectively causes loss of cell membrane integrity,mitochondrial metabolic arrest, depletion of RNA, DNA lysis andaggregation, and cell fragmentation and cell loss. The apoptotic processleads to typical ultrastructural progressive changes of membranousdisruption and swelling, progressively deepening nuclear invaginationswith eventual membranous bleb formations and cell death andfragmentation into apoptotic bodies. Histologically, typical apoptoticchanges with positive immunohistochemical staining of markers forapoptosis are found throughout the injected areas for up to severalweeks after treatment.

FT has been extensively tested in patients with BPH. The compound andplacebo controls have been administered by the transrectal route in over1700 procedures in 9 human clinical trials. In these large long-termclinical trials in men with BPH, FT was administered in a concentrationof 0.25 mg/ml (2.5 mg of FT—amounting to administration to about 15-20%of the gland by volume). See, e.g., Shore, et al., “The potential forNX-1207 in benign prostatic hyperplasia: an update for clinicians,” TherAdv. Chronic Dis., 2(6), pp. 377-383 (2011). It therefore is preferredthat compositions comprising FT include at least 2.5 mg of FT, and canbe administered in amounts of up to 25 mg of FT in a singleadministration. In another embodiment, FT is administered in an amountwithin the range of from about 2.5 mg to about 20 mg, or from about 2.5mg to about 15 mg. In an embodiment, FT is administered in an amount of15 mg.

The following examples are provided to illustrate the presentembodiments. It should be understood, however, that the embodiments arenot to be limited to the specific conditions or details described inthese examples. Throughout the specification, any and all references toa publicly available document, including a U.S. patent, are specificallyincorporated by reference. In particular, the embodiments expresslyincorporate by reference the examples contained in U.S. Pat. Nos.6,924,266; 7,241,738; 7,317,077; 7,408,021; 7,745,572; 8,067,378;8,293,703; 8,569,446; and 8,716,247, and U.S. Patent ApplicationPublication Nos. 2017/0360885; 2017/0020957; 2016/0361380; and2016/0215031, each of which reveal that certain peptides specifiedtherein are effective agents for causing cell death in vivo in normalrodent muscle tissue, subcutaneous connective tissue, dermis and othertissue.

EXAMPLES

In a series of clinical studies a total of 146 men with low gradeprostate cancer (Gleason grade 6) were treated in the following manner.Patients were randomized and treated with a transrectal intraprostaticsingle injection of a composition comprising 2.5 mg FT (n=49), or acomposition comprising 15 mg FT (n=48), or were subject to controlactive surveillance (n=49). After the first follow-up biopsy at 45 dayspost-randomization, 18 patients in the control active surveillance groupcrossed over to a single administration of a composition in which 10patients received 2.5 mg FT, and 8 patients received 15 mg FT. Patientswere followed for 5 years including biopsies at baseline, 45 days, 18,36, and 48 months, and urological evaluations with PSA every 6 months.Patients with Gleason grade increase or who elected surgical orradiotherapeutic intervention exited the study and were still includedin the data analysis. Percentage of normal biopsies in the baselinefocus quadrant and median tumor grade and volume were assessed;progression was measured by clinical and pathological outcomes includingGleason grade, and Gleason grade primary pattern, in the entire prostatesampled as well as for the treated prostate lobe. Interventionsassociated with Gleason grade increase as well as total incidence forany conventional surgical intervention (e.g., surgery, radiotherapy,and/or chemotherapy) was assessed.

Example 1

This example evaluated the percentage of patients who exhibited greaterthan one new focus (i.e., went from unifocal to multifocal) in theentire prostate, in which the new focus had an increase in Gleason gradetotal score (“new multifocal with Gleason upgrade”). The data in thetables below for the times of follow up represent the percentage ofpatients who exhibited progression or worsening. The results are shownin Table 3 below:

TABLE 3 Time from treatment and % improvement Treatment <=18 mos % Imp.<=36 mos % Imp. <=48 mos % Imp. FT 15 mg 5.7 (p = 0.025) 79.5 9.1 (p =0.13) 79.2 20 (p = 0.039) 65 FT 2.5 mg 13.5 51.4 23.1 47.2 50 12.4Pooled FT 9.7 (p = 0.044) 65.1 16.7 (P = 0.027) 61.9 35.5 37.8 Control27.8 43.8 57.1

The results from example 1 reveal that the embodiments can reduce thepercentage of patients who had entire prostate new multifocal cancerwith Gleason upgrade, when compared to active surveillance controls,when measured at least 18 months after treatment, by an amount of fromabout 40% to about 100%, or from about 50% to about 90%, or from about50% to about 80%, or any value therebetween. The method of theembodiments can reduce the percentage of patients who had entireprostate new multifocal cancer with Gleason upgrade, when compared toactive surveillance controls, when measured at least 36 months aftertreatment, by an amount of from about 30% to about 90%, or from about45% to about 90%, or from about 47% to about 80%, or any valuetherebetween. The method of the embodiments can reduce the percentage ofpatients who had entire prostate new multifocal cancer with Gleasonupgrade, when compared to active surveillance controls, when measured atleast 48 months after treatment, by an amount of from about 5% to about85%, or from about 10% to about 70%, or from about 12% to about 65%, orany value therebetween.

Example 2

This example evaluated the percentage of patients who exhibited greaterthan one new focus (i.e., went from unifocal to multifocal) in theentire prostate, in which the new focus had an increase in Gleasonprimary pattern (“new multifocal with increase Gleason primarypattern”). The results are shown in Table 4 below:

TABLE 4 Time from treatment and % improvement Treatment <=18 mos % Imp.<=36 mos % Imp. <=48 mos % Imp. FT 15 mg 2.9 82.6 4.5 82 13.3 62.7 FT2.5 mg  0 (p = 0.17) 100  0 (p = 0.074) 100  0 (p = 0.009) 100 Pooled FT1.4 (p = 0.049) 91.6 2.1 (p = 0.0031) 91.6 6.5 (p = 0.012) 81.8 Control16.7 25 35.7

The results from example 2 reveal that the embodiments can reduce thepercentage of patients who had entire prostate new multifocal cancerwith increase in Gleason primary pattern, when compared to activesurveillance controls, when measured at least 18 months after treatment,by an amount of from about 40% to about 100%, or from about 70% to about100%, or from about 80% to about 100%, or any value therebetween. Themethod of the embodiments can reduce the percentage of patients who hadentire prostate multifocal cancer with increase in Gleason primarypattern, when compared to active surveillance controls, when measured atleast 36 months after treatment, by an amount of from about 30% to about100%, or from about 70% to about 100%, or from about 80% to about 100%,or any value therebetween. The method of the embodiments can reduce thepercentage of patients who had entire prostate multifocal cancer withincrease in Gleason grade primary pattern, when compared to activesurveillance controls, when measured at least 48 months after treatment,by an amount of from 20% to about 100%, or from about 50% to about 100%,or from about 60% to about 100%, or any value therebetween.

Example 3

This example evaluated the percentage of patients who exhibited greaterthan one new focus (i.e., went from unifocal to multifocal) in thehemi-prostate in which the initial unifocal tumor was treated, in whichthe new focus had an increase in Gleason grade total score(“hemi-prostate multifocal cancer with Gleason upgrade”). The resultsare shown in Table 5 below:

TABLE 5 Time from treatment and % improvement Treatment <=18 mos % Imp.<=36 mos % Imp. <=48 mos % Imp. FT 15 mg 5.7 69.7 9.1 (p = 0.049) 74.513.3 70.8 FT 2.5 mg 8.6 54.26 16.7 53.2 42.9 5.7 Pooled FT 7.1 62.2 13.0(p = 0.0549) 63.6 27.6 39.3 Control 18.8 35.7 45.5

The results from example 3 reveal that the embodiments can reduce thepercentage of patients who had multifocal prostate cancer in the lobe(or hemi-prostate) in which the foci were initially treated with Gleasonupgrade (patients with >=1 new focus in treated lobe or hemi-prostatewith increased Gleason grade), when compared to active surveillancecontrols, when measured at least 18 months after treatment, by an amountof from about 40% to about 100%, or from about 50% to about 90%, or fromabout 60% to about 80%, or any value therebetween. The method of theembodiments can reduce the percentage of patients who had hemi-prostatemultifocal cancer with Gleason upgrade, when compared to activesurveillance controls, when measured at least 36 months after treatment,by an amount of from about 50% to about 90%, or from about 50% to about85%, or from about 50% to about 80%, or any value therebetween. Themethod of the embodiments can reduce the percentage of patients who hadhemi-prostate multifocal cancer with Gleason upgrade, when compared toactive surveillance controls, when measured at least 48 months aftertreatment, by an amount of from about 15% to about 80%, or from about15% to about 75%, or from about 16% to about 72%, or any valuetherebetween.

Example 4

This example evaluated the percentage of patients who exhibited greaterthan one new focus (i.e., went from unifocal to multifocal) in thehemi-prostate in which the initial unifocal tumor was treated, in whichthe new focus had an increase in Gleason grade primary pattern 4(“hemi-prostate multifocal cancer with increase in Gleason primarypattern”). The results are shown in Table 6 below:

TABLE 6 Time from treatment and % improvement Treatment <=18 mos % Imp.<=36 mos % Imp. <=48 mos % Imp. FT 15 mg  2.9 76.8  4.5 79  6.7 75.5 FT2.5 mg   0 (p = 0.033) 100  0 (p = 0.018) 100  0 (p = 0.037) 100 PooledFT 1.4 (p = 0.03) 88.8 2.2 (p = 0.011) 89.7 3.4 (p = 0.025) 87.5 Control12.5 21.4 27.3

The results from example 4 reveal that the embodiments can reduce thepercentage of patients who had hemi-prostate multifocal cancer withincrease in Gleason primary pattern, when compared to activesurveillance controls, when measured at least 18 months after treatment,by an amount of from about 50% to about 100%, or from about 70% to about100%, or from about 75% to about 100%, or any value therebetween. Themethod of the embodiments can reduce the percentage of patients who hadhemi-prostate multifocal cancer with increase in Gleason primarypattern, when compared to active surveillance controls, when measured atleast 36 months after treatment, by an amount of from about 45% to about100%, or from about 70% to about 100%, or from about 75% to about 100%,or any value therebetween. The method of the embodiments can reduce thepercentage of patients who had hemi-prostate multifocal cancer withincrease in Gleason primary pattern, when compared to activesurveillance controls, when measured at least 48 months after treatment,by an amount of from 45% to about 100%, or from about 60% to about 100%,or from about 70% to about 100%, or any value therebetween.

Example 5

This example evaluated the percentage of patients who had conventionalcancer treatment (e.g., surgery, radiotherapy, and/or chemotherapy), andwho exhibited greater than one new focus (i.e., went from unifocal tomultifocal) in the entire prostate, in which the new focus had anincrease in Gleason grade total score (“cancer treatment with newmultifocal with Gleason upgrade”). The results are shown in Table 7below:

TABLE 7 Time from treatment and % improvement Treatment <=18 mos % Imp.<=36 mos % Imp. <=48 mos % Imp. FT 15 mg 5.1 (p = 0.02) 80.6 7.1 (p =0.17) 79.9 8.3 (p = 0.003) 83.4 FT 2.5 mg  9.8 62.7 16.7 52.7 26 48Pooled FT 7.5 (p = 0.02) 71.5 12.1 (p = 0.026) 65.7  17 (p = 0.009) 66Control 26.3 35.3 50

The results from example 5 reveal that the embodiments can reduce thepercentage of patients who had cancer treatment with new multifocal withGleason upgrade in the entire prostate, when compared to activesurveillance controls, when measured at least 18 months after treatment,by an amount of from about 50% to about 100%, or from about 55% to about90%, or from about 60% to about 80%, or any value therebetween. Themethod of the embodiments can reduce the percentage of patients who hadcancer treatment with new multifocal with Gleason upgrade in the entireprostate, when compared to active surveillance controls, when measuredat least 36 months after treatment, by an amount of from about 40% toabout 90%, or from about 45% to about 90%, or from about 50% to about80%, or any value therebetween. The method of the embodiments can reducethe percentage of patients who had cancer treatment with new multifocalwith Gleason upgrade in the entire prostate, when compared to activesurveillance controls, when measured at least 48 months after treatment,by an amount of from about 35% to about 85%, or from about 40% to about85%, or from about 48% to about 84%, or any value therebetween.

Example 6

This example evaluated the percentage of patients had conventionalcancer treatment (e.g., surgery, radiotherapy, and/or chemotherapy), andwho exhibited greater than one new focus (i.e., went from unifocal tomultifocal) in the entire prostate, in which the new focus had anincrease in Gleason grade primary pattern ≥4 (“cancer treatment with newmultifocal and an increase in Gleason grade primary pattern”). Theresults are shown in Table 8 below:

TABLE 8 Time from treatment and % improvement Treatment <=18 mos % Imp.<=36 mos % Imp. <=48 mos % Imp. FT 15 mg 2.6 (p = 0.06) 83.5 3.6 (p =0.04)  84.7 4.2 (p = 0.019) 86.6 FT 2.5 mg   0 (p = 0.001) 100  0 (p =0.006) 100   0 (p = 0.0040) 100 Pooled FT 1.3 (p = 0.04) 91.8 1.7 (p =0.002) 92.8 2.1 (p = 0.001) 93.3 Control 15.8 23.5 31.3

The results from example 6 reveal that the embodiments can reduce thepercentage of patients having cancer treatment with new multifocal andan increase in Gleason grade primary pattern in the entire prostate,when compared to active surveillance controls, when measured at least 18months after treatment, by an amount of from about 70% to about 100%, orfrom about 75% to about 100%, or from about 80% to about 100%, or anyvalue therebetween. The method of the embodiments can reduce thepercentage of patients who had cancer treatment with new multifocal andan increase in Gleason grade primary pattern in the entire prostate,when compared to active surveillance controls, when measured at least 36months after treatment, by an amount of from about 60% to about 100%, orfrom about 70% to about 100%, or from about 80% to about 100%, or anyvalue therebetween. The method of the embodiments can reduce thepercentage of patients who had cancer treatment with new multifocal andan increase in Gleason grade primary pattern in the entire prostate,when compared to active surveillance controls, when measured at least 48months after treatment, by an amount of from 50% to about 100%, or fromabout 70% to about 100%, or from about 80% to about 100%, or any valuetherebetween.

Example 7

This example evaluated the percentage of patients who had conventionalcancer treatment (e.g., surgery, radiotherapy, and/or chemotherapy), andwho and who had multifocal prostate cancer in the lobe (orhemi-prostate) in which the foci were initially treated with Gleasonupgrade (cancer treatment with new multifocal with increased Gleasongrade in the hemi-prostate). The results are shown in Table 9 below:

TABLE 9 Time from treatment and % improvement Treatment <=18 mos % Imp.<=36 mos % Imp. <=48 mos % Imp. FT 15 mg 5.1 67.7 7.1 69.8 8.3 73.5 FT2.5 mg 4.9 69 10 57.4 17.4 44.4 Pooled FT 5 68.4 8.6 63.4 12.8 59.1Control 15.8 23.5 31.3

The results from example 7 reveal that the embodiments can reduce thepercentage of patients who had cancer treatment with new multifocalcancer with Gleason upgrade in the hemi-prostate, when compared toactive surveillance controls, when measured at least 18 months aftertreatment, by an amount of from about 40% to about 100%, or from about50% to about 90%, or from about 60% to about 75%, or any valuetherebetween. The method of the embodiments can reduce the percentage ofpatients who had cancer treatment with new multifocal with increasedGleason grade in the hemi-prostate, when compared to active surveillancecontrols, when measured at least 36 months after treatment, by an amountof from about 50% to about 90%, or from about 55% to about 75%, or fromabout 50% to about 80%, or any value therebetween. The method of theembodiments can reduce the percentage of patients who had cancertreatment with new multifocal with increased Gleason grade in thehemi-prostate, when compared to active surveillance controls, whenmeasured at least 48 months after treatment, by an amount of from about15% to about 80%, or from about 35% to about 75%, or from about 40% toabout 75%, or any value therebetween.

Example 8

This example evaluated the percentage of patients having conventionalcancer treatment (surgery, radiotherapy, or chemotherapy), and havingmultifocal prostate cancer in the lobe (or hemi-prostate) in which thefoci were initially treated with Gleason primary pattern (cancertreatment with new multifocal and an increase in Gleason grade primarypattern in the hemi-prostate). The results are shown in Table 10 below:

TABLE 10 Time from treatment and % improvement Treatment <=18 mos % Imp.<=36 mos % Imp. <=48 mos % Imp. FT 15 mg  2.6 75.2  3.6 79.5  4.2 77.7FT 2.5 mg  0 (p = 0.025) 100   0 (p = 0.017) 100  0 (p = 0.031) 100Pooled FT 1.3 (p = 0.034) 87.6 1.7 (p = 0.01) 90.3 2.1 (p = 0.019) 88.8Control 10.5 17.6 18.8

The results from example 8 reveal that the embodiments can reduce theembodiments can reduce the percentage of patients who had cancertreatment with new multifocal cancer with an increase in Gleason gradeprimary pattern in the hemi-prostate, when compared to activesurveillance controls, when measured at least 18 months after treatment,by an amount of from about 65% to about 100%, or from about 70% to about100%, or from about 75% to about 100%, or any value therebetween. Themethod of the embodiments can reduce the percentage of patients who hadcancer treatment with new multifocal and an increase in Gleason gradeprimary pattern in the hemi-prostate, when compared to activesurveillance controls, when measured at least 36 months after treatment,by an amount of from about 65% to about 100%, or from about 70% to about100%, or from about 75% to about 100%, or any value therebetween. Themethod of the embodiments can reduce the percentage of patients who hadcancer treatment with new multifocal and an increase in Gleason gradeprimary pattern in the hemi-prostate, when compared to activesurveillance controls, when measured at least 48 months after treatment,by an amount of from 60% to about 100%, or from about 65% to about 100%,or from about 75% to about 100%, or any value therebetween.

The results from the foregoing examples illustrate the unexpectedlysuperior effect of pharmaceutically active ingredients, andspecifically, FT in reducing multi-focal cancer incidence, reducingmulti-focal cancer grade, and reducing multi-focal cancer progression(worsening) in the entire prostate or in the hemi-prostate where theinitial low grade tumor was located and treated. It will be apparent tothose skilled in the art that various modifications and variations canbe made in the methods and compositions of the present embodimentswithout departing from the spirit or scope of the embodiments.

What is claimed is:
 1. A method of reducing multifocal prostate cancerincidence, reducing multifocal prostate cancer grade, and reducingmultifocal prostate cancer progression in a mammal having a unifocalprostate cancer tumor comprising: administering to the unifocal prostatecancer a composition comprising a therapeutically effective amount offexapotide triflutate (FT); and wherein administering FT to only the lowgrade unifocal prostate cancer tumor reduces multifocal prostate cancerincidence, reduces multifocal prostate cancer grade, and reducesmultifocal prostate cancer progression in the entire prostate.
 2. Themethod of claim 1, wherein the composition is administered by injectiondirectly to the unifocal cancer tumor.
 3. The method of claim 1, whereinthe unifocal cancer tumor is a cancerous tumor of a multifocal cancerselected from the group consisting of prostate cancer, squamous cellhead and neck carcinoma (HNSCC), cutaneous squamous cell carcinoma (skinSCC), breast cancer, adenocarcinoma and SCC of lung, esophageal cancer,gastric cancer, colon cancer, bladder cancer, cervical cancer, melanoma,brain cancer, pancreatic cancer, ovarian cancer, bone marrow cancer, andleukemia.
 4. The method of claim 1, wherein the method reduces newmultifocal prostate cancer with Gleason grade increase in the entireprostate by: a) an amount of from about 40% to about 100%, when comparedto active surveillance, when measured at least 18 months aftertreatment; b) an amount of from about 30% to about 90%, when compared toactive surveillance, when measured at least 36 months aftertreatment;and c) an amount of from about 5% to about 85%, when compared to activesurveillance, when measured at least 48 months after treatment.
 5. Themethod of claim 1, wherein the method reduces new multifocal prostatecancer with an increase in Gleason grade primary pattern in the entireprostate by: a) an amount of from about 40% to about 100%, when comparedto active surveillance, when measured at least 18 months aftertreatment; b) an amount of from about 30% to about 100%, when comparedto active surveillance, when measured at least 36 months aftertreatment; and c) an amount of from about 20% to about 100%, whencompared to active surveillance, when measured at least 48 months aftertreatment.
 6. The method of claim 5, wherein the method reducesconventional cancer treatment and new multifocal prostate cancer withGleason grade increase in the entire prostate by: a) an amount of fromabout 50% to about 100%, when compared to active surveillance, whenmeasured at least 18 months after treatment; b) an amount of from about40% to about 90%, when compared to active surveillance, when measured atleast 36 months aftertreatment; and c) an amount of from about 35% toabout 85%, when compared to active surveillance, when measured at least48 months after treatment.
 7. The method of claim 1, wherein the methodreduces conventional cancer treatment and new multifocal prostate cancerwith an increase in Gleason grade primary pattern in the entire prostateby: a) an amount of from about 70% to about 100%, when compared toactive surveillance, when measured at least 18 months aftertreatment; b)an amount of from about 60% to about 100%, when compared to activesurveillance, when measured at least 36 months aftertreatment; and c) anamount of from about 50% to about 100%, when compared to activesurveillance, when measured at least 48 months after treatment.
 8. Themethod of claim 1, wherein the method reduces conventional cancertreatment and new multifocal prostate cancer with an increase in Gleasongrade primary pattern in the prostate lobe where the low grade unifocalprostate cancer focus was located by: a) an amount of from about 65% toabout 100%, when compared to active surveillance, when measured at least18 months after treatment; b) an amount of from about 65% to about 100%,when compared to active surveillance, when measured at least 36 monthsaftertreatment; and c) an amount of from about 60% to about 100%, whencompared to active surveillance, when measured at least 48 months aftertreatment.
 9. The method of claim 1, wherein the therapeuticallyeffective amount of FT is within the range of from about 2.5 mg to about20 mg.
 10. The method of claim 1, wherein the therapeutically effectiveamount of FT is within the range of from about 2.5 mg to about 15 mg.11. The method of claim 1, wherein the therapeutically effective amountof FT is 15 mg.